International Journal of Pure and Applied Zoology Volume 4, Issue 2, pp: 142-148, 2016 ISSN (Print) : 2320-9577 ISSN (Online): 2320-9585 *Corresponding author e-mail: sadyzola@uol.com.br 142 http://www.ijpaz.com Research Article EFFECTS OF THE EXPOSURE TO ATRAZINE AND GLYPHOSATE THROUGHOUT INCUBATION ON BONE DEVELOPMENT OF PODOCNEMIS EXPANSA (TESTUDINES, PODOCNEMIDIDAE) Juliana dos Santos Mendonça1, Lucélia Gonçalves Vieira1, Sady Alexis Chavauty Valdes2*, André Luiz Quagliatto Santos1 1 Wild Animal Research and Teaching Laboratory (LAPAS), Universidade Federal de Uberlândia (UFU), Uberlândia, MG, Brazil 2 Centro Universitário de Patos de Minas (UNIPAM), Patos de Minas, MG, Brazil INTRODUCTION Technological progress enabled the evolution of several areas related to food production, health, and agriculture. However, these improvements are intimately connected with the use of chemical compounds that still have unknown properties (Oga, 2003). Nowadays, pesticides are the most important environmental contaminants from human activities that cause severe problems to living organisms. Given the characteristics of these compounds and their persistence in the biosphere, this kind of pollutant may affect natural communities, causing impacts and deleterious effects at tissue and molecular levels in animals (Bueno-Guimarães et al., 2001; Berti et al., 2009). The use of herbicides in weed control has been recognized all over the world as a useful agricultural practice. However, its indiscriminate use may impact non-target organisms, mainly aquatic organisms (Nwani et al., 2010). Brazil is considered the largest consumer of pesticides in the world (ANDEF, 2012), and herbicides, mainly atrazine and glyphosate, are the most used class of pesticides in the country (Cox, 2001; IBAMA, 2010b). Atrazine (2-chloro-4-ethylamine-6-isopropylamine-s- triazine) is more commonly used in rural areas, mainly in corn, sorghum, and sugarcane crops. Although it is classifed as moderately toxic for aquatic species, this herbicide is one of the most common contaminants detected in streams, rivers, lakes, dams, and underground waters (Battaglin et al., 2003; Scrubner et al., 2005; Battaglin et al., 2008). On the other hand, glyphosate (N-(phosphonomethyl) glycine) may be the most important herbicide ever developed (World Health Organization, 1994). Due to its low persistence in the soil and biodegradability, glyphosate is applied repeatedly in weed control of several crops. As, glyphosate sales represent 76% of the total trade of herbicides in Brazil (IBAMA, 2009), large amounts of this pesticide may reach non-target organisms (Mitchell et al., 1987; Servizi et al., 1987). In 2006, the legal regulations of the herbicide glyphosate started to be reviewed in Brazil. Today, this review request is under analysis, according to the resolution RDC no. 10/2008 (ANVISA, 2014). After eight years of study, the Genetics and Environmental Mutagenesis Research Group (GEMA) made up by researchers from the Universidade Nacional de Río Cuarto (UNRC), issued a report in which they linked the use of glyphosate with genetic changes that may lead to spontaneous miscarriage, fetal malformation, and cancer (Torres et al., 2006). However, the greatest obstacle ABSTRACT Brazil is considered to be one of the largest consumers of pesticides in the world, and herbicides represent the most commonly used class of these products. Atrazine and glyphosate, the most important herbicides, contaminate the waters of several Brazilian rivers and are involved in organ and bone malformations in different species. Among the possible target organisms, reptiles may be indirectly affected by pesticide use once their natural habitats are rivers and streams. Therefore, the objective of this study was to assess the possible effects of exposure to the herbicides atrazine and glyphosate in bone ontogeny of Podocnemis expansa. Eggs were artifcially incubated in sand moistened with water contaminated with atrazine at concentrations equal to 0, 2, 20, or 200 μg/L, and glyphosate at 65, 650 or 6,500 μg/L. In the control group, the substrate was moistened with distilled water. Two eggs were collected from each incubator every ten days until hatching. For the analysis of bone development, soft tissues were diaphanized, and bones and cartilages were stained with Alizarin red S and Alcian blue, respectively. Specimens were analyzed by stereomicroscopy. Morphological characteristics of the cartilages and bones of the embryos were compared with descriptions of normal ontogeny available in the literature for P. expansa embryos. No abnormalities were observed in bone ontogeny of any of the experimental groups. Keywords: Ecotoxicology; Skeleton; Herbicides; Reptiles; South American River Turtle Article History: Received 08 th January 2016; Accepted 12 th February 2016; Published 18 th February 2016